Kit busting: Enhanced Chemiluminescence

ashendel at ashendel at
Fri Nov 12 01:12:35 EST 1993

I read and have since lost a reference to a paper that described the use of 
phenols for stabilizing the chemiluminescence generated by the action of 
HRP+hydrogenperoxide on luminol.  I believe that this may provide a clue to 
the nature of the magic (proprietary) ingredient Amersham includes in its 
ECL kit solutions.  As the rest of the reagents are quite inexpensive 
(H2O2, luminol, buffer), I would like to see if I can obtain equivalent 
results with some of the phenols indicated in that paper to make a do-it-
yourself ECL(tm)-like reagent mix (for detection of peroxidase on western 
blots) at a fraction of the $215 kit price charged by Amersham.  I am aware 
of the NEN BLAST (tm) kit.  Also, an old Worthington reference on HRP 
indicaes that it tolerates up to 170 mM phenol in the assay.  

Has anyone done this successfully?  What is a good phenol to use and at 
what concentration?  What is the citation for this paper?

My apologies if this was discussed previouly.  Thanks in advance,

Curt Ashendel
Dept. Medicinal Chemistry, Purdue University

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